Dissertations / Theses on the topic 'Canine babesiosis'

Canine babesiosis typically causes a haemolytic anemia and results in hypoxia and sepsis, which can eventually result in multiple organ dysfunction. Human patients with severe injury or disease such as shock, sepsis and malaria often have persistent hyperlactataemia, and there is a correlation between blood lactate and survival rate. There are various similarities between human malaria and canine babesiosis, eg. anaemia, renal failure, cerebral forms, coagulopathy, hepatopathy, pulmonary oedema, and shock. In severe malaria, lactate levels in blood rise in direct proportion to the severity of the disease. Venous lactate concentrations measured at 4 hours after admission appears to be the best prognostic indicator in severe malaria. In dogs blood lactate has been shown to be of prognostic value in patients with gastric dilatationvolvulus and in dogs admitted to intensive care units. Blood lactate has also been shown to be of prognostic value in equine colic. Blood lactate was determined in ninety dogs with naturally occurring canine babesiosis. Forty-five dogs (50%) presented with hyperlactataemia (blood lactate > 2.5mmol/L) and 20 (22.2%) with hypoglycaemia (blood glucose < 3.3 mmol/L). Measurements significantly associated with mortality were hypoglycaemia on admission, blood lactate > 5mmol/L on admission, blood lactate > 2.5 mmol/L at 8, 16 and 24 hours after admission, and increase or < 50% decrease in blood lactate within 8 and 16 hours after admission. Blood lactate persistently > 4.4 mmol/L indicated a very poor prognosis. The study concluded that serial blood lactate measurements are useful in predicting survival in dogs with severe and complicated canine babesiosis.
Dissertation (MMed Vet (Med))--University of Pretoria, 2004.
Companion Animal Clinical Studies
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Canine babesiosis is an emerging infectious disease in the United States (US). An epidemic of Babesia gibsoni infections in the US was identified. An association between dog breed and B. gibsoni infections was detected. Babesia gibsoni-infected dogs were more likely to be American pit bull terriers and B. canis vogeli infected dogs were more likely to be greyhounds. An association between a recent dog bite and B. gibsoni infection was detected, implicating direct dog-to-dog transmission as a route of infection in the US. Several genes from canine Babesia spp. were characterized, including 18S ribosomal RNA (rRNA), internal transcribed spacer regions (ITS), cytochrome B (cytB), and rhoptry-associated protein-1 (RAP-1). These genetic data were used to develop a sensitive and specific diagnostic semi-nested polymerase chain reaction (PCR) test for canine babesiosis. Using this assay, a novel large Babesia organism was identified in a blood sample obtained from a clinical patient. Molecular and phylogenetic characterization of this large Babesia spp. determined that it was most closely related to B. bigemina. Lastly, an atovaquone and azithromycin drug combination was shown to be the first treatment to clear canine B. gibsoni infections.

The pathology of canine cerebral babesiosis was examined at the gross, histological and ultrastructural levels. Gross lesions could be categorised as either global or regional. Congestive brain swelling , diffuse cerebral congestion and diffuse cerebral pallor were classified as global lesions. Multifocal haemorrhage and malacia were classified as regional lesions. Oedema was inconsistently present and could be either focal or diffuse. The majority of histological changes were observed in both cerebral babesiosis and control cases. Regional lesions were unique to cerebral babesiosis and had specific histological features. Highly localised endothelial injury was the primary lesion. Early lesions were multifocal and strictly associated with the microvasculature. Intermediate lesions, with perivascular haemorrhage and neutrophil infiltration, were suggestive of reperfusion injury. Advanced lesions were locally extensive and similar in appearance to haemorrhagic infarction. It is likely that the pathogenesis of regional lesions is by a process of microvascular infarction, as venous thrombosis could not be demonstrated. Ultrastructural evidence for adherent contact between erythrocytes and capillary endothelium was demonstrated. Endothelial cell necrosis occurred early in the development of lesions, before neuronal and glial injury. It is postulated that endothelial injury is the primary event in the development of regional lesions and secondary lesions develop as a consequence of microvascular infarction.
Die patologie van die serebrale vorm van bosluiskoors in honde is ondersoek. Die letsels is makroskopies, histologies en elektronmikroskopies beskryf. Letsels kon makroskopies in twee groepe verdeel word: Globale letsels en gelokaliseerde letsels. Kongestiewe brein swelling, diffuse serebrale kongestie en serebrale anemie kom voor as globale letsels in serebrale babesiose. Multifokale bloeding en nekrose kom voor as gelokaliseerde letsels. Edeem was nie konsekwent teenwoordig nie, en was algemeen of verspreid. Die meeste algemene histologiese veranderinge was in beide serebrale en kontrole gevalle teenwoordig. Gelokaliseerde letsels waarin spesifieke hisotpatologiese veranderinge voorgekom het, was kenmerkend van serebrale babesiose. Die primere letsel is hoogs gelokaliseerde beskadiging van endoteelselle. Beskadiging van die kapillere bloedvate ontstaan vroeg in die ontwikkeling van letsels. Verdere ontwikkeling van die letsel word gekenmerk deur peri-vaskulere bloeding en neutrofiel infiltrasie wat aanduidend is van reperfusie beskadiging. Volontwikkelde letsels is plaaslik-ekstensief en het die voorkoms van hemoragiese infarkte Dit is waarskynlik dat mikrovaskulere infarksie 'n rol speel in die patogenese van die letsels, aangesien veneuse trombose nie ontstaan nie. Noue kontak tussen rooibloedselle en kapillere endoteel is elektronmikroskopies bevestig. Endoteelselnekrose ontstaan voordat tekens van beskadiging geidentifiseer kan word in neurone of gliaselle. Dit blyk dat kapillere endoteelselbeskadiging die primere letsel by die ontstaan van gelokaliseerde lese Is is, en dat sekondere lesels ontwikkel as gevolg van mikrovaskulere infarksie.
Dissertation (MSc)--University of Pretoria, 2000.
Paraclinical Sciences
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Electrocardiographic (ECG) changes have never been reported in canine babesiosis. Based on the metabolic, electrolyte, and myocardial alterations described for the disease, such changes are to be expected. The purpose of this study was to describe ECG changes in canine babesiosis, and to correlate those changes to clinical severity, outcome and cardiac histopathological changes. Four groups of dogs with babesiosis were studied: mild to moderate anaemia (n=40), severe anaemia (n=35), concurrent immune-mediated haemolytic anaemia (n=18) and complicated (n=28). Lead II ECG was recorded at admission for 1 minute in all dogs, and repeated after 24 hours in admitted dogs (groups II – IV). Six lead ECG was recorded in 88 dogs. Full necropsy was performed between 30-60 minutes after death on 16 dogs (5 died on arrival, 11 had ECG recording). Gross cardiac pathology was recorded and histopathology of myocardial sections from ventricles, atria, apex and interventricular septa was evaluated, using a scoring system for haemorrhages, necrosis, inflammatory infiltrate and fibrin microthrombi. The following ECG changes were recorded: sinoatrial (7%) and atrioventricular blocks (4%), ventricular premature complexes (7%), low R-amplitude (23%), prominent Q (33%), axis deviations (40%), prolonged QRS (32%), ST depression and coving (28%), large T (42%), and notched R (28%). Differences between groups were minor and inconsistent. Gross pathological changes were pericardial effusion (25%) and subepicardial (56%) and subendocardial haemorrhages (63%). Histological changes were haemorrhages (69%), necrosis (50%), inflammation (63%) and fibrin microthrombi (75%). The only correlation between pathology and ECG was low R-amplitude and pericardial effusion. There was a significantly higher prevalence of sinus bradycardia and irregular sinus rhythm in the non-survivors. Both ECG and pathological changes were non-specific, but there were similarities to the pattern of changes that have been described for myocarditis and myocardial ischaemia. Antiarrhythmic treatment was only required in 1 dog. Thus, the clinical application of the ECG changes found in this study was limited. It was concluded that the heart suffers from the same pathological processes described in other organs in canine babesiosis, namely inflammation and hypoxia. Cardiovascular management, if necessary, should be based on functional monitoring rather than ECG.
Dissertation (MMedVet (Med))--University of Pretoria, 2001.
Companion Animal Clinical Studies
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Babesiosis, caused by Babesia rossi, is a common cause of morbidity and mortality of dogs in South Africa. Canine babesiosis can be classified either as uncomplicated or complicated based on the degree of anaemia and the severity of the presenting clinical signs.1,2 In uncomplicated babesiosis, the clinical signs are mostly attributable to the degree of the anaemia, whereas in complicated babesiosis the disease process is characterised by additional organ involvement.3,4 One of the most common haematological hallmarks of canine babesiosis, caused by B. rossi, is thrombocytopenia, which is not associated with clinical haemorrhage despite very low platelet counts that would normally cause inability to maintain normal primary haemostatic function.5 The aim of this study was to describe the thromboelastographic findings in uncomplicated canine babesiosis and compare them with those of normal, healthy control dogs. We hypothesised that these dogs would have a normal to hypercoagulable haemostatic capacity, despite the severe thrombocytopenia, and that this could be detected with thromboelastography (TEG), which has previously been shown to correlate well with clinical signs of haemorrhage in dogs.6 This was a prospective, cross sectional, observational study that included 20 client-owned dogs, diagnosed with uncomplicated canine babesiosis at the Onderstepoort Veterinary Academic Hospital (OVAH). Infection with B. rossi was confirmed by polymerase chain reaction (PCR) and reverse line blot (RLB) hybridisation assay. Blood samples were collected at the time of diagnosis. A group of 10 healthy control dogs were included for comparison. Antithrombin activity (AT) was measured using an automated spectrophotometric analyser (Cobas Integra 400, Roche, South Africa). D-dimer was measured using an immunometric flow-through principle (D-dimer Single test, Nycocard Reader II, Medinor A/S). Prothrombin time (PT), activated partial thromboplastin time (aPTT) and fibrinogen assays were performed on the ST art® 4 analyser (Diagnostica Stago, Roche, South Africa). TEG analysis was performed using the TEG® 5000 Thromboelastograph® Haemostasis System (Haemoscope, Pro-Gen Diagnostics (Pty) Ltd, South Africa). A complete blood count was performed on the ADVIA 2120 (Siemens, South Africa). The results of the babesiosis and control groups were compared using the Mann-Whitney U test or the Students t-test based on normality. The normality assumption for distribution of the variables in the data was evaluated using the Shapiro-Wilk test. The statistical significance was set at p<0.01. The mean haematocrit (Ht) and median platelet count was significantly lower in the babesiosis group than the controls (0.29 vs. 0.50 L/L; p<0.01 and 22.0 vs. 374.5 x 109/l; p<0.01, respectively). There was no significant difference in any of the TEG parameters between the babesiosis group and the controls. The medians for the various TEG parameters in the babesiosis group versus the controls were; R: 5.5 vs. 4.4 min (p=0.05); K: 2.5 vs. 2.0 min (p=0.08); angle: 58.3 vs. 61.1 degrees (p=0.35); MA: 47.0 vs. 57.0 mm (p=0.02); G: 4.9 vs. 6.7 dyn/cm2 (p=0.02); LY30: 0.00 vs. 0.6% (p=0.20); and LY60: 0.00 vs. 3.0% (p=0.014). The median fibrinogen concentration was significantly higher in the babesiosis group than in the control group; 5.8 g/L (5.0 – 7.0) vs. 2.9 g/L (2.5 – 3.3); (p<0.01). The mean AT activity was significantly lower in the babesiosis group than in the control group; 102.6 mg/dl (89.9 – 112.8) vs. 127.8 mg/dl (110.6 – 134.8); (p<0.01). The median D-dimer concentration was not significantly different in the babesiosis group compared to the control group; 0.3 mg/L (0.1 – 0.4) vs. 0.1 mg/L (0.1 – 0.2); (p=0.016). Median PT was not significantly different in the babesiosis group compared to the control group; 6.5 sec (6.4 – 7.2) vs. 6.8 sec (6.6 – 7.5); (p=0.14). Median aPTT was significantly prolonged in the babesiosis group compared to the control group; 13.6 sec (12.4 – 14.5) vs. 11.5 sec (10.7 – 12.2); (p<0.01). Despite the severe thrombocytopenia, dogs suffering from uncomplicated babesiosis did not have clinical signs of haemorrhage. The thromboelastograms of the babesiosis group were normal to hypercoagulable and thus correlated well with the clinical phenotype. Copyright
Dissertation (MSc)--University of Pretoria, 2011.
Companion Animal Clinical Studies
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This thesis showed that dogs with natural infection with B. canis had both renal and cardiac dysfunction, both of which can be classified as complications of babesiosis and would thus necessitate supportive therapy. This thesis demonstrated that RTE celluria, proteinuria, and variable enzymuria and azotaemia occur in dogs with babesiosis. However, these were all minimal changes and all could be consistent with hypoxia, reduced GFR, or reduced cardiac output This thesis showed that dogs with naturally occurring babesiosis had significant urine met-haemoglobin with no evidence of blood met-haemoglobin. The possibility would be that the urinary methaemoglobin was either produced in the kidney or possibly by oxidation of haemoglobin to met-haemoglobin in the bladder. It has been shown experimentally that met-haemoglobin can be toxic. The combination of reduced GFR, anaemic hypoxia, and met-haemoglobin can all act synergistically to cause renal damage. Renal haemodynamics are also much more likely to be abnormal when cardiac dysfunction is present Reduced renal blood flow and glomerular filtration rate are evidence of redistribution of blood flow that commonly occurs in early heart failure. An important finding in this thesis was that dogs with babesiosis had lower serum sodium than control dogs but there was no difference between mild, severe, or complicated cases of babesiosis. In addition, dogs with babesiosis had a lower fractional clearance of sodium than Clinically healthy control dogs, which can be interpreted as sodium retention by the kidneys. This sodium retention would also result in water retention , which will result in an expansion of the plasma volume. In the past heart lesions in canine babesiosis were regarded as rare complications, with the majority of lesions being reported as incidental findings at post-mortem examination of complicated babesiosis cases. This thesis has demonstrated that cardiac lesions to be common in canine babesiosis. This thesis showed that that ECG changes in babesiosis were similar to the pattern described for myocarditis and myocardial ischaemia, and together with the histopathological findings indicated that the heart suffers from the same pathological processes described in other organs in canine babesiosis, namely inflammation and hypoxia. As the clinical application of the ECG changes found in this thesis was limited, cardiovascular assessment should be based on functional monitoring rather than ECG. Using cardiac troponin as a marker of myocardial injury, this thesis showed that myocardial cell injury occurs with canine babesiosis. Cardiac troponins, especially troponin I, are sensitive markers of myocardial injury in canine babesiosis, and the magnitude of elevation of plasma troponin I concentrations appears to be proportional to the severity of the disease. ECG changes and serum cardiac troponin were correlated with histopathology. On cardiac histopathology from dogs that succumbed to babesiosis, haemorrhage, necrosis, inflammation and fibrin microthrombi in the myocardium were documented, all of which would have resulted in ECG changes and elevations in cardiac troponin. Myocardial infarction causes left ventricular failure, which will result in hypotension and an expansion of the plasma volume due to homeostatic mechanisms. This thesis showed that dogs with babesiosis had hypoalbuminaemia, which may be because of intravascular volume dilution due to fluid retention. In disease hypoalbuminaemia can occur as a negative acute-phase protein. In the light of the cardiac changes, hyponatraemia, and hypotension, a probable cause would be fluid retention due to myocardial disease. This thesis showed that dogs with babesiosis had left ventricular lesions, which can result in systolic heart failure.
Thesis (PhD)--University of Pretoria, 2005.
Veterinary Tropical Diseases
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Abnormal carbohydrate metabolism is a commonly encountered feature of malaria in people, and similar derangements have been detected in veterinary patients with canine babesiosis. Glucose, the major metabolic fuel source, is a key resource in critically ill patients as they mount an immunological response to infection and inflammation. The ability of the individual to effectively mobilise, distribute and utilise glucose is a major determinant of morbidity and mortality. Hypoglycaemia has been identified as a life threatening metabolic complication in almost 20% of severely ill dogs suffering from babesiosis due to Babesia rossi infection. Insulin and glucagon are the primary hormones involved in glucose homeostasis. Insulin lowers blood glucose concentration by facilitating cellular uptake and utilisation of glucose. Hyperinsulinaemia as a result of inappropriate insulin secretion may precipitate hypoglycaemia, and has been identif ed as a cause of hypoglycaemia in human and murine malaria. A similar phenomenon may exist in canine babesiosis. This prospective, cross-sectional, observational study, including 94 dogs with naturally acquired virulent babesiosis, sought to investigate and characterise the relationship between blood glucose concentrations and insulin concentrations in cases of canine babesiosis. Pre-treatment jugular blood samples were collected for simultaneous determination of plasma glucose and insulin concentrations. Animals were retrospectively divided into three groups: hypoglycaemic (plasma glucose concentration < 3.3 mmol/L; n=16), normoglycaemic (3.3-5.5 mmol/L; n=62), and hyperglycaemic (> 5.5 mmol/L; n=16). The median plasma insulin concentrations (IQR in parentheses) for the hypoglycaemic, normoglycaemic and hyperglycaemic groups were 10.7 pmol/L (10.7-18.8 pmol/L), 10.7 pmol/L (10.7-29.53 pmol/L; i.e below the detection limit of the assay), and 21.7 pmol/L (10.7-45.74 pmol/L), respectively. Statistical analysis revealed no significant difference in insulin concentration between the three groups. These results suggest that insulin secretion was appropriately suppressed in these dogs. Only two dogs had elevated insulin concentrations, one of which was hypoglycaemic. The median time since last meal (available for 87 dogs) was 24 hours (IQR 2-4 days), constituting a significant period of illness-induced starvation. We conclude that hyperinsulinaemia is not a cause of hypoglycaemia in virulent canine babesiosis. It is speculated that prolonged fasting due to disease-induced anorexia, in addition to increased glucose consumption, depletion of hepatic glycogen stores, and hepatic dysfunction with impaired gluconeogenesis, may play important roles in the pathophysiology of hypoglycaemia in canine babesiosis.
Dissertation (MMedVet)--University of Pretoria, 2010.
Companion Animal Clinical Studies
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Made available in DSpace on 2016-08-16T18:18:41Z (GMT). No. of bitstreams: 1 Tese Andressa Almeida Santana.pdf: 1466363 bytes, checksum: ae0eefc75fb09cd9f96594f4e5656ed5 (MD5) Previous issue date: 2011-07-29
FUNDAÇÃO DE AMPARO À PESQUISA E AO DESENVOLVIMENTO CIENTIFICO E TECNOLÓGICO DO MARANHÃO
Canine babesiosis and canine visceral leishmaniasis (CVL) are vector borne diseases, where dogs exert a play as reservoir or source for arthropods responsible by the transmission of these protozoosis. Babesia canis vogeli, is transmited by tick Rhipicephalus sanguineus while Leishmania infantum (sin. Leishmania chagasi) is transmited by sand fly (Lutzomyia longipalpis). The results showed that CVL remains endemic in São Luís Municipality. Despite that the coinfection between Leishmania and Babesia was low considering that both diseases are endemic in this tropical area. Beside that was observed that Yorkshire terrier presented higher predisposition to acquire the infection by B. canis vogeli. A remarkable result was the occurrence of ocular lesions associated to L. infantum infection.
A babesiose canina e a leishmaniose visceral canina são doenças transmitidas por vetores, sendo os cães competentes reservatórios e fonte alimentar dos artrópodes envolvidos. Babesia canis vogeli, é um parasita intraeritrocitário transmitido pelo carrapato Rhipicephalus sanguineus. Leishmania infantum (sin. Leishmania chagasi) transmitidos a mamíferos pela picada dos flebotomíneos (Lutzomyia longipalpis), infectando macrófagos do Sistema Fagocítico Mononuclear do hospedeiro. O presente trabalho é divido em capítulos e os resultados encontrados mostraram que a leishmaniose visceral canina ainda é endêmica no município de São Luís, e que apesar disso a taxa de coinfecção com Babesia foi baixa. Também foi observado que a raça Yorkshire terrier, dentre as raças estudadas, apresentou maior predisposição para contrair a infecção por B. canis vogeli. Outro resultado significativo foi a ocorrência de lesões oculares associadas à infecção por L. infantum.

Acute phase proteins (APP) are ideal biomarkers for inflammation due to their stability, relative ease of assay and apparent relation between their concentration and the extent of the insult to tissue. C-reactive protein (CRP) is a positive major APP in dogs and can be used as a predictive marker for risk of disease and to monitor the response to treatment. Increased concentrations in certain diseases are associated with poor outcome. This cross-sectional, observational study of 75 dogs naturally infected with Babesia rossi, a cause of virulent canine babesiosis, was designed to examine the association of CRP concentration at admission and the magnitude of CRP change 24 hours after admission with outcome. Dogs were excluded if there was evidence of concurrent inflammatory diseases at the time of admission, infection with subtypes other than B. rossi, concurrent Ehrlichia canis infections or euthanasia for reasons other than poor prognosis. Diagnosis was confirmed by polymerase chain reaction and reverse line blot. CRP concentrations were determined by an automated human CRP Turbidometric Immunoassay (TIA), previously validated for use in dogs (Bayer CRP TIA, Newbury, UK), on serum samples collected by jugular venipuncture on admission, prior to any therapy, and thereafter daily until discharge or death. There was no significant difference in admission CRP concentration between survivors (n = 57; median = 97.4 mg/l; mean ± SD = 107.5 ± 49.5), and non-survivors (n = 11; median = 101.4 mg/l; mean ± SD = 122.1 ± 64.6) (p = 0.39). After elimination of non-significant predictors, a multiple exact logistic regression model for predicting mortality contained glucose and CRP. Mortality was associated with decreased glucose levels (p = 0.0002) and increased CRP levels (p = 0.045) on admission. Multiple regression analysis failed to show a significant relationship between admission CRP concentration and number of days of hospitalization in the survivors, adjusting for age and sex (p = 0.65). No significance was found in the relationship between the magnitude of change in CRP concentration 24 hours after admission, and the number of days of hospitalization in survivors, (p = 0.34). Using an admission CRP concentration cut-off of 60 mg/l, survival proportions between the two groups were no different (p = 0.34) and when applied to the group of dogs that survived, it was not associated with length of hospitalization (p = 0.25). In corroboration with previous reports glucose was identified as a major prognostic marker for mortality, but additionally the pro-inflammatory marker CRP was identified as a significant co-prognosticator. Copyright
Dissertation (MMedVet)--University of Pretoria, 2009.
Companion Animal Clinical Studies
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In order to investigate the increased serum urea: creatinine ratio encountered in canine babesiosis in South Africa, packed cell volume (PCV), serum haemoglobin, serum total bilirubin, plasma ammonia, serum urea, serum creatinine, calculated serum urea: creatinine ratio, serum cystatin-C, total serum protein (TSP) and urine analysis were performed in this study. The model consisted of 28 canine babesiosis patients, 25 in group 1, the anaemic group with PCV < 20%, and 3 in group 3, the haemoconcentrated group with PCV > 50%. 13 canine babesiosis negative dogs made up group 2, the control group. Unpublished human malaria data from 363 patients was also analysed and the urea: creatinine ratio was calculated in order to determine whether elevated serum urea: creatinine ratios also occur in this disease. The serum urea: creatinine ratio was > 20 in 23/25 anaemic patients, 3/13 control patients and 2/3 haemoconcentrated patients. The mean and median serum urea: creatinine ratio for the anaemic babesia group was 46.32 and 41.36, for the haemoconcentrated babesia group 25.10 and 20.85, for the malaria patients 20.64 and 18.75, and for the control group 15.91 and 15.36 respectively. Mean and median serum urea: creatinine ratio was significantly elevated and the distribution varied significantly between the anaemic babesiosis group compared with the control group (p=0.000003). Serum urea: creatinine ratio was elevated in canine babesiosis patients, especially when they are anaemic. The serum urea: creatinine ratio was also found to be elevated in the human malaria patients. Serum urea was found to be elevated in 17/25 anaemic patients, 0/13 control patients and 3/3 haemoconcentrated patients. Mean and median values for serum urea were 12.83 mmol/l and 11.8 mmol/l and 5.09 mmol/l and 4.3 mmol/l for the anaemic group and the control group respectively. Mean and median serum urea was significantly elevated and the distribution varied significantly between the anaemic group compared with the control group (p=0.00003). Serum creatinine was elevated in 0/25 anaemic patients 0/13 control patients and 2/3 haemoconcentrated patients. Mean and median values for serum creatinine were 72.08 mmol/l and 67 mmol/l, and 78.69 mmol/l and 75 mmol/l for the anaemic group and the control group respectively. Mean, median and distribution of serum creatinine did not vary significantly between the control group and the anaemic babesia group (p=0.274488). Serum cystatin-C was elevated in 0/25 anaemic patients, 0/13 control patients and 2/3 haemoconcentrated patients. The two haemoconcentrated patients with elevated serum urea and serum creatinine also had elevated serum cystatin-C. One of these two patients was available for post mortem and had significant renal lesions on histopathology. These two patients also had a urine specific gravity (SG) < 1.030 (1.020 for both cases). Mean and median values for serum cystatin-C were 0.71 mg/l and 0.7 mg/l, and 0.67 mg/l and 0.7 mg/l for the anaemic group and the control group respectively. Mean, median and distribution of serum cystatin-C did not vary significantly between the control group and the anaemic babesia group, (p=0.450172). Using the Spearman-rank test for correlation coefficients serum creatinine but not serum urea had a significant positive correlation with serum cystatin-C; and serum urea had a significant positive correlation with the serum urea: creatinine ratio. Log serum creatinine was significantly correlated to log serum cystatin-C, (r2=0.52, p<0.001). Log serum urea was also significantly correlated to log serum cystatin-C, (r2=0.14, p=0.024), however, this correlation was relatively poor compared to that of serum creatinine with serum cystatin-C. Log serum urea: creatinine ratio was not significantly correlated with log serum cystatin-C, (r2=0.014, p=0.485). Given its resistance to interference from the elevated plasma and serum pigments found in canine babesiosis, an elevated serum cystatin-C was taken to indicate significant renal disease. Therefore we can deduce that the increased serum urea: creatinine ratio encountered in canine babesiosis is a result of disproportionately elevated serum urea concentrations, which is of non-renal origin. A similar event appears to be present in human falciparum malaria patients. The cause of this phenomenon in either disease is unknown; but various forms of prerenal azotaemia, hyperureagenesis and laboratory perturbations may play a role. Serum creatinine was a better measure of significant renal disease than serum urea in this study. There also appeared to be no added benefit of measuring serum cystatin-C. Mean and median values for PCV were 11.64% and 11%, and 46.08% and 48% for the anaemic group and the control group respectively. As expected the mean and median PCV was significantly lower and the distribution varied significantly between the control group and the anaemic babesia group (p=0.000001). The mean and median values for PCV in the haemoconcentrated group were 54.33% and 54% respectively. Serum haemoglobin was > 1.6 g/l in 8/25 anaemic patients, 0/13 control patients and 3/3 haemoconcentrated patients. Mean and median values for serum haemoglobin were 1.72 g/l and 1.3 g/l, and 0.88 g/l and 0.8 g/l for the anaemic group and the control group respectively. Mean and median serum haemoglobin was significantly elevated and distribution of serum haemoglobin varied significantly between the control group and the anaemic babesia group (p=0.002364). Serum total bilirubin was > 15 mmol/l in 11/25 anaemic patients, 0/3 haemoconcentrated patients and 0/13 control patients. Mean and median values for serum total bilirubin were 22.42 mmol/l and 10.6 mmol/l, and 10.63 mmol/l and 11.0 mmol/l for the anaemic group and the control group respectively. The median and the distribution of serum total bilirubin did not vary significantly between the control group and the anaemic babesia group (p=0.355888). TSP was elevated in 5/25 anaemic patients, was below the normal laboratory reference range in 5/25 anaemic patients, was elevated in 1/13 control patients, and was normal in all the haemoconcentrated cases. Mean and median values for TSP were 58.60 g/l and 54 g/l, and 60.92 g/l and 60.0 g/l for the anaemic group and the control group respectively. The mean, median and distribution of TSP did not vary significantly between the control group and the anaemic babesia group (p=0.130661).Plasma ammonia was elevated in 16/25 anaemic patients, 0/13 control patients and 2/3 haemoconcentrated patients. Mean and median values for plasma ammonia were 78.77 mmol/l and 53.8 mol/l, and 23.18 mmol/l and 21.5 mmol/l for the anaemic group and the control group respectively. Mean and median plasma ammonia was significantly elevated and the distribution of plasma ammonia varied significantly between the control group and the anaemic babesia group (p=0.009562). Using the Spearman-rank test for correlation coefficients PCV had a significant negative correlation with plasma ammonia, serum urea and serum urea: creatinine ratio. Serum haemoglobin and serum bilirubin had a significant positive correlation with plasma ammonia, serum urea and serum urea: creatinine ratio. Plasma ammonia also had a significant positive correlation with serum urea and serum urea: creatinine ratio. These correlations may be explained by substrate loading as a result of haemolysis. I speculate that various protein by-products of haemolysis may cause the hyperammonaemia encountered via deamination of these proteins. This in turn may lead to ureagenesis, the physiological process of ammonia clearance. This ureagenesis may be excessive and therefore may account for the elevated serum urea encountered in patients with normal serum creatinine and serum cystatin-C concentrations. Alternatively the presence of an elevated serum haemoglobin concentration with its high intrinsic absorptive capacity in the 300 nm to 500 nm wave length range, may positively bias the measurement of serum urea and plasma ammonia concentrations. Serum urea is measured at 340 nm and plasma ammonia is measured at 365 nm. Thus a non-physiological process could also explain the above correlations. Urine analysis of the canine babesiosis patients showed mild evidence of renal disease as portrayed by proteinuria, renal tubular epithelium (RTE) celluria and granular casts. However, these findings, as well as the presence of an elevated serum urea, haemoglobinaemia and haemoglobinuria, bilirubinaemia and bilirubinuria, were present in both patients with mild renal disease and patients with overt renal disease. Therefore the benefit of single serum urea concentrations and random urine sediment analysis in canine babesiosis is questioned. Serial urine analysis monitoring was not investigated. This study concluded that serum urea is often elevated due to non-renal factors in canine babesiosis patients. This causes an elevated serum urea: creatinine ratio in this disease, with a similar event appearing to occur in human malaria patients. The cause of these elevated ratios remains undetermined, but is likely to be as a result of hyperureagenesis or laboratory error. The measurement of serum creatinine, urine SG and hourly urine production is advocated to detect significant renal disease in these patients. There appeared to be no added benefit derived from the measurement of serum cystatin-C concentrations in canine babesiosis patients in this preliminary study.
Dissertation (MMedVet (Medicine))--University of Pretoria, 2005.
Companion Animal Clinical Studies
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Blood transfusion forms a mainstay of the treatment of a variety of illnesses, and is lifesaving. Nonetheless, it is not without its risks and drawbacks. Blood transfusion is a cornerstone in the treatment of canine babesiosis. The development of blood alternatives has received attention in recent times. Blood alternatives offer much of what natural blood does but without many of the associated drawbacks. These include disease transmission, transfusion reactions, poor in vitro and in vivo shelf-life and special storage and administration requirements. One product, Oxyglobin®, is the first commercially available, veterinary-licensed, haemoglobin-based oxygen carrying solution (HBOCS). Although licenced for use in canine babesiosis, this colloidal “Oxygen Bridge” has never been evaluated against the gold standard of therapy, isovolumic packed red blood cell transfusion (pRBCT). This investigation was conducted to evaluate important aspects the equivalence of these two treatments in a field situation of naturally-infected dogs. Given the cost of HBOCS, they are unlikely to be commonly used by the practicing veterinarian in the treatment of canine babesiosis. Nonetheless, similarities in efficacy would bolster the case for and further research into blood substitutes of this and other classes, and may open the way to evaluation of HBOCS for falciparum malarial anaemia, a disease similar in many respects to canine babesiosis.
Dissertation (MMedVet)--University of Pretoria, 2007.
Companion Animal Clinical Studies
unrestricted

Babesiosis is a disease caused by parasites of Babesia species that is spread through ticks. Babesiosis can affect humans and many other mammals all over the world. In the United States, babesiosis is mainly caused by Babesia microti with additional species of Babesia infecting dogs. Dogs have long been known to be a good indicator species for human tick-borne infection due to the shared spaces they have with humans and their tendency to explore and pick up diseases in the environment. This study used a group of hunting dogs to determine the presence of Babesia infection. The goal was to determine a predictive model for human infection. Infection was defined as having a positive antibody test or molecular test for Babesia species in blood samples. Blood samples were taken at two time points, tested, and compared. Statistical methods were used to analyze the results of the tests and compare them with variables such as region, gender, age, and other diseases the dogs were exposed to. Of 214 dogs, 56 had Babesia infections, with a prevalence rate of 26.2%. At the first time point the model showed age and infection with Anaplasma platys as being significant. At the 2nd time point, 29 dogs were lost to follow up, leaving a total of 185 dogs sampled with 63 testing positive for Babesia spp. The rate for the 2nd time point was 34.1%. No variables were found to be significant in the model for the 2nd time point. Despite differences in Babesia that infect humans and dogs, the hunting dog cohort reflects a model that validates the environmental exposures, coinfections, and demographic variables that affect transmission of the pathogen. Furthermore, findings of this study cast doubt on the ability of Ixodes scapularis to act as capable vector for canine-infecting Babesia species.

Šunų babeziozė – tai liga, greitai plintanti visame pasaulyje, kurią sukelia pirmuonys, priklausantys Babesia genčiai. Ligos sukėlėjus platina Dermocentor, Rhipicephalus, Haemaphysalis genties erkės. Lietuvoje paplitę Dermacentor reticulatus erkės - pagrindiniai Babesia canis vektoriai. B. canis patenka į raudonąsias kraujo ląsteles (eritrocitus) ir jas suardo. Kasmet sergamumas šunų babezioze visoje Europoje bei Lietuvoje stipriai auga. Darbo tikslas buvo įvertinti šunų užsikrėtimą Babesia canis patogenais ir identifikuoti Babesia canis porūšius ir padermes, panaudojant molekulinius tyrimo metodus ir filogenetinę sekų analizę. Šiame darbe buvo tiriami 20 šunų, sergančių babezioze,( pagal babeziozei būdingą klinikinę išraišką) kraujo mėginiai, kurie buvo surinkti iš Kauno apskrities Veterinarijos klinikų. Taikant molekulinį PGR (polimerazės grandininės reakcijos) metodą, 17 iš šių mėginių buvo identifikuota Babesia canis canis. Atlikus palyginamąją Babesia canis canis 18S rRNR geno sekų analizę, pasitelkus Gen Bank duomenis, buvo nustatyta, kad Lietuvoje sekvenuotos sekos nuo užsikrėtusių šunų buvo identiškos tarpusavyje ir turėjo 91-100% panašumą su sekomis, identifikuotomis sergančiuose šunyse ir D. reticulatus erkėse kitose Europos šalyse. Identifikuotos sekos priklausė vienam iš genotipų, paplitusių Europoje, kuris pasižymi didesniu virulentiškumu. Europos šalyse nustatyta 10 B. canis canis sekų variantų, identifikuotų pagal 18S rRNR geno sekų analizę.
Canine babesiosis is a disease, quickly spreading worldwide, which is caused by the protozoa belonging to the genus Babesia. The disease agents are transmitted by ticks of the genera Dermocentor, Rhipicephalus, and Haemaphysalis. Dermacentor reticulatus ticks, which are widely spread in Lithuania, are the main vectors of Babesia canis. B. canis infects red blood cells (erythrocytes) and destroys them. Every year the morbidity of Babesia canis is intensely growing in the whole of Europe and Lithuania. The aim of thesis was to evaluate the canine infection with Babesia canis pathogens and identify species and subspecies of Babesia canis, using molecular methods of analysis and phylogenetic sequence. The research focused on 20 dogs infected with Babesia canis (in accordance with the clinical expression characteristic of babesiosis); the blood samples were collected in the veterinary clinics in the Kaunas region. Applying the molecular PCR (Polymerase chain reaction) method, 17 of the samples were identified as cases of Babesia canis canis. On the basis of the contrastive gene sequencing analysis of Babesia canis canis 18S rRNR and the data of Gen Bank it was determined that in Lithuania the sequential sequences of infected dogs were identical and by 91-100 percent resembled the sequences identified in infected dogs and D. reticulatus ticks in other European countries. The identified sequences belonged to one of the genotypes which is widely spread in Europe and is... [to full text]

Tema: babezioze sergančių šunų gydymo ypatybės. Tyrimai buvo atliekami Lietuvos Sveikatos Mokslų Universiteto, Veterinarijos Akademijos, Dr. L. Kriaučeliūno smulkiųjų gyvūnų veterinarijos klinikoje. Darbo apimtis 46 puslapiai, jame yra 6 lentelės, 16 paveikslų, panaudoti 72 literatūros šaltiniai. Tyrimų tikslas – išanalizuoti babezioze sergančių šunų gydymo ypatybes, įvertinti galimas komplikacijas bei jų įtaką prognozei. Tyrimų uždaviniai: 1) Atlikti detalią babezioze sergančių šunų klinkinių požymių analizę. 2) Išanalizuoti ir pateikti diagnostinių priemonių, naudojamų šunų babeziozės diagnozavimui bei diferenciacijai, spektrą. 3) Atlikti vaistų ir kitų šunų babeziozės gydymui naudojamų metodų analizę. Įvertinti naudojamų antiparazitinių preparatų privalumus ir trūkumus. Tyrimų metodika: 1) Į kliniką atvestų šunų, kuriems įtariama babeziozė, šeimininkų prašyta nurodyti šunų amžių, veislę, laikymo ir šėrimo sąlygas, pastebėtus pirmuosius ligos požymius, kada jie pasireiškė, bei ar rado paskutiniu metu į šunį įsisiurbusių erkių. 2) Buvo atlikti periferinio kraujo tyrimai, kraujo morfologiniu bei biocheminiu rodikliu tyrimai. 3) Statistinė duomenų analizė atlikta skaičiuokle „Excel 2010“. Rezultatai ir išvados: Šunims, sirgusiems babezioze, dažniausiai pasireiškė apatija ir vangumas (100 proc. atvejų), karščiavimas (69 proc. atvejų), blogas apetitas (58,6 proc. atvejų), splenomegalija (41,4 proc. atvejų), galinių kojų silpnumas (37,9 proc. atvejų), mažakraujystė (34,5 proc... [toliau žr. visą tekstą]
The topic: treatment specifics of canine babesiosis. Researches were carried out in Lithuanian University of Health Sciences, Veterinary Academy, Dr. L. Kriaučeliūno veterinary clinic for small animals. Paper volume 46 Pages, it contains 6 tables, 16 images, used 72 references. The research objective – to analyze the treatment specifics for dogs with babesiosis, evaluate the potential complications and their impact on prognosis. Reseach tasks: 1) Perform a detailed analysis of clinical signs in dogs with babesiosis. 2) Analyze and provide diagnostic tools used to diagnose dog babesiosis and differentiation spectrum. 3) Perform analysis on drugs and other treatment methods used in the therapy for dogs with babesiosis. Research methodologies: 1) Dogs owners were asked to indicate their dog's age, breed, housing and feeding conditions, the first signs of the disease which they noticed, when they occurred, and if they found ticks on their dogs recently. 2) Peripheral blood research, blood morphological and biochemical indicators tests were made. 3) The statistical analysis was carried out using calculator in „Excel 2010 “. Results and conclusions: Dogs with a history of babesiosis, commonly observed lethargy and sluggishness (100 percent of cases), fever (69 percent of cases)., loss of appetite (58.6 percent of cases)., splenomegaly (41.4 percent of cases)., rear leg weakness (37.9 percent of cases)., anemia (34.5 percent of cases). Babesiosis had little effect on anemia - red... [to full text]

Darbo tikslas – surinkti ir išanalizuoti šunų sergančių babezioze atvejus, organų bei jų funkcijų sutrikimus komplikuotoje babeziozės formoje smulkių gyvūnų veterinarijos klinikoje, Klaipėdoje. Darbo uždaviniai: nustatyti kokio amžiaus, veislės bei lyties šunys dažniausiai serga komplikuota babeziozės forma; nustatyti kurio sezono metu daugiausiai pasireiškia komplikuota babeziozės forma; ištirti ir įvertinti biocheminių kraujo tyrimų rezultatus; nustatyti ar sergamumas komplikuota babeziozės forma priklauso nuo sudėtinio organų bei jų funkcijų sutrikimo, bei jo įtaką mirtingumui; nustatyti kokių organų sistemų bei jų funkcijų sutrikimai vyrauja dažniausiai ir ar šių organų sistemų veikla daro įtaką mirtingumui. Tyrimo objektas: šunys sergantys komplikuota babeziozės forma. Tyrime taikyti diagnostikos analizės metodai, statistika, grafika, palyginimas, mokslinės literatūros analizė, sisteminis požiūris, matematiniai metodai, koreliacijos metodas. Išvados: 1. Iš 32 šunų kuriems buvo nustatyta komplikuota babeziozės infekcija dažniausiai (41%) sirgo 1-5 metų šunų amžiaus grupė, dažniausiai sirgo mišrūnai 13 (39,4%), rečiau škotų terjerai 3 (9%), sergančių patinų užregistruota 62,5 %, patelių 37,5%. 2. Didžiausi ligos protrūkiai pasireiškė pavasario (47%) bei rudens (37%) sezono metu 3. Analizuojant kraujo biocheminius parametrus buvo nustatyta, kad babeziozės kraujo biocheminiai parametrai buvo nukrypę nuo normos ribų: šlapalo koncentracija buvo virš normos ribų 3,1 – 8,5... [toliau žr. visą tekstą]
This study was designed to collect and analyze the dog with babesiosis cases, organ dysfunction, and their a complicated form of babesiosis of small animal veterinary clinic in Klaipeda. The research objective formulated in the following tasks: identify what age, breed and sex dogs usually suffer from a complicated form of babesiosis; set a season mainly occurs in the form of complicated babesiosis; investigate and evaluate the biochemical blood tests; determine whether the incidence of babesiosis complicated form depends on a complex of organs and their functioning, and its impact on mortality; determine what organ systems and their functions, disorders usually prevail and that the organ systems function affects mortality. The object of study: Dogs with complicated babesiosis form. In the research apply the methods of diagnostic analysis, statistics, graphic, comparison, the scientific literary analysis, systematic approach, mathematical methods of treatment, correlation method, survey method. Conclusions: 1. Of the 32 dogs who were with complicated babesiosis infection in most cases (41 %) had 1-5 years of age group of dogs , mostly bastards had 13 (39.4 % ), followed by the Scottish terrier , 3 (9 %) , with 62.5 % of males recorded and females 37.5 % 2. The largest outbreaks occurred in the spring (47 %) and autumn (37 %) during the season 3. The analysis of blood biochemical parameters were found to Babesia in blood biochemical parameters were abnormal values: blood urea... [to full text]

O relatório de estágio de Mestrado Integrado em Medicina Veterinária, aqui apresentado, encontra-se dividido em duas secções. A primeira corresponde à descrição das atividades médico-veterinárias desenvolvidas ao longo do estágio curricular e a segunda secção corresponde à revisão bibliográfica de babesiose canina. O estágio foi realizado no Hospital Animal do Sul, situado em Faro e abrangeu a área de medicina interna e cirurgia de animais de companhia. A revisão bibliográfica é seguida do relato e discussão de um caso clínico de babesiose não complicada, acompanhado durante o período de estágio. A babesiose canina faz parte das doenças transmitidas por vetores com grande importância mundial e, apesar de ainda não estarem descritos casos humanos provocados por espécies de Babesia responsáveis por doença em cães, existem outros fatores que devem ser tidos em conta na análise da babesiose canina como uma futura potencial zoonose; Abstract Canine Babesiosis This report of Integrated Masters Degree in Veterinary Medicine, here presented, is divided into two sections. The first one corresponds to the description of medicalveterinary activities developed throughout the internship and the second one corresponds to a literature review on canine babesiosis. The internship was perfomed in Hospital Animal do Sul, in Faro and has as main areas of interest internal medicine and surgery of domestic animals. The literature review is followed by a presentation and discussion of a case-report of a uncomplicated babesiosis case, followed during the internship period. Canine babesiosis is one of vector-borne diseases of worldwide importance and, although human cases caused by species of Babesia responsible for disease in dogs, haven’t been reported, there are other factors that should be considered in the analysis of canine babesiosis as a future potential zoonosis.

Tikslas: išanalizuoti įvairių veiksnių įtaką šunų babeziozės pasireiškimui, plitimui, atsižvelgiant į šuns lytį, amžių ir veislę, naudojant literatūros šaltinius bei asmeninius tyrimus. Duomenys buvo surinkti iš " Jakovo veterinarijos centras", smulkių gyvūnų klinikos "Pas Filą " ir kraujo tyrimai buvo renkami ir atliekami Veterinarijos ir diagnostikos centre. Iš viso surinkome 1195 ligos atvejų, atsižvelgiant į lytį, amžių bei veislę. Anamnezė buvo surinkta iš 117 šunų, kraujo mėginiai buvo imami iš 31 sergančio šuns, kraujo morfologiniams tyrimams. Išanalizavus esamus duomenis, pastebėjome, kad dažniausiai babezioze serga tam tikros šunų veislės: mišrūnai, vokiečių aviganiai, labradoro retriveriai ir sibiro haskiai. Atsižvelgiant į lytį, tai daugiausiai babezioze sirgo patinai ir žymiai rečiau patelės. Babezeliozė dažniausiai buvo diagnozuojama jauniems šunims, 1 iki 2 metų, bet sunkiausia klinika buvo pasireiškusi šuniukams iki 1 metų. Dažniausiai pasireiškianti klinikinė simptomatika: apatija, anoreksija, kliniškai išreikšta anemija, pakitusi kūno temperatūra, tamsus šlapimas, vėmimas, NS sutrikimai, viduriavimas, kliniškai išreikšta gelta. Analizuojant morfologinius kraujo rodiklių pokyčius, nustatėmė, kad visais atvejais buvo pasireiškusi trombocitopenija, sumažėjęs trombokrito kiekis. Pastebėjome, kad ligai būdingas sezoniškumas. Stipriausias babeziozės bumas pasireiškia pavasarį – balandžio, gegužės, birželio mėnesiais ir kiek silpnesnis – rudenį (rugsėjį, spalį). ... [toliau žr. visą tekstą]
Goal: to find out a variety of factors influence the occurrence of canine babesiosis, development, according to dog sex, age and breed, using known literature and personal research. The data were collected from „Jakovo veterinary center“, small animal clinic „pas Filą“ and the blood tests were collected and conducted at Veterinaty and Diagnostic center. We collected 1195 disease cases according to sex, age and breed. Anamnesis were collected from 117 dogs, morphological studies of blood – 31. According to the studies, we found out, that there are certain breeds are more susceptible to babesiosis: mongrels, german shepherds, labrador retrievers and siberian huskies. Also found, that babesiosis more prone males than females. Dog babesiosis were usually diagnosed in young dogs, 1 to 2 years, but the hardest clinic gets puppies up to 1 year. The most frequent clinical manifestations in patients with babesiosis : apathy, anorexia, symptomatic anemia, change in body temperature, dark urine, vomiting, NS disorders, diarrhea, clinical jaundice. Morphological characteristics of the blood we found that in all cases, is characterized by thrombocytopenia, a decrease thrombocyte. The seasonal variation in the number of cases has been reported at all Veterinary clinics, with a higher incidence in the spring months (April, May) and peaking in autumn (September, November).

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Universidade Estadual Paulista (UNESP)
Com o objetivo avaliar a epidemiologia e a característica molecular da infecção de cães de áreas rurais por Babesia spp., amostras de sangue de 150 animais foram coletadas e analisadas por esfregaço sangüíneo e por PCR. Além disso, os cães foram examinados para investigar possível infestação por carrapatos. Pelo esfregaço sangüíneo de sangue capilar (ponta de orelha) foram detectados três (2%) cães positivos e pela PCR para Babesia spp. 12 (8%) deles foram positivos, com bandas visualizadas em 450 pb. Foram encontrados R. sanguineus e Amblyomma spp. em 36 (24%) dos 150 cães. As espécies de Amblyomma detectadas foram A. cajennense (9/36 - 25%), A. ovale (8/36 - 22,2%) e A. aureolatum (1/36 - 2,7%). Um espécime de Amblyomma (1/36 - 2,7%) não pôde ser identificado especificamente pois se encontrava no estágio de ninfa. Não houve uma associação positiva entre o encontro de R. sanguineus e infecção por B. canis. O seqüenciamento de uma amostra positiva demonstrou identidade com B. canis vogeli já caracterizada no Brasil.
In order to study the parasitologic and morphological characteristics of Babesia spp. infection in dogs from rural areas, 150 animals were examined by blood smears and by PCR. Besides, the dogs were examined to investigate ticks infestation. By the blood smear of capillary blood (ear tip) examination, three (2%) were detected positive and by PCR for Babesia spp.12 (8%) were positive, with bands visualized in 450 bp. Rhipicephalus sanguineus and Amblyomma spp. were found in 36 (24%) of the 150 dogs. The found species of Amblyomma were A. cajennense (9/36 - 25%), A. ovale (8/36 - 22.2%) and A. aureolatum (1/36 - 2.7%). A specimen of Amblyomma (1/36 - 2.7%) was found in the nymph stage and could not be identified. There was not a positive association between the encounter of R. sanguineus and the infection for B. canis. The sequencing of two positive samples demonstrated close identity with B. canis vogeli already characterized in Brazil.

Orientador: Lucia Helena O'Dwyer de Oliveira
Banca: Teresa Cristina Goulart de Oliveira-Sequeira
Banca: Vamilton Álvares Santarém
Resumo: Com o objetivo avaliar a epidemiologia e a característica molecular da infecção de cães de áreas rurais por Babesia spp., amostras de sangue de 150 animais foram coletadas e analisadas por esfregaço sangüíneo e por PCR. Além disso, os cães foram examinados para investigar possível infestação por carrapatos. Pelo esfregaço sangüíneo de sangue capilar (ponta de orelha) foram detectados três (2%) cães positivos e pela PCR para Babesia spp. 12 (8%) deles foram positivos, com bandas visualizadas em 450 pb. Foram encontrados R. sanguineus e Amblyomma spp. em 36 (24%) dos 150 cães. As espécies de Amblyomma detectadas foram A. cajennense (9/36 - 25%), A. ovale (8/36 - 22,2%) e A. aureolatum (1/36 - 2,7%). Um espécime de Amblyomma (1/36 - 2,7%) não pôde ser identificado especificamente pois se encontrava no estágio de ninfa. Não houve uma associação positiva entre o encontro de R. sanguineus e infecção por B. canis. O seqüenciamento de uma amostra positiva demonstrou identidade com B. canis vogeli já caracterizada no Brasil.
Abstract: In order to study the parasitologic and morphological characteristics of Babesia spp. infection in dogs from rural areas, 150 animals were examined by blood smears and by PCR. Besides, the dogs were examined to investigate ticks infestation. By the blood smear of capillary blood (ear tip) examination, three (2%) were detected positive and by PCR for Babesia spp.12 (8%) were positive, with bands visualized in 450 bp. Rhipicephalus sanguineus and Amblyomma spp. were found in 36 (24%) of the 150 dogs. The found species of Amblyomma were A. cajennense (9/36 - 25%), A. ovale (8/36 - 22.2%) and A. aureolatum (1/36 - 2.7%). A specimen of Amblyomma (1/36 - 2.7%) was found in the nymph stage and could not be identified. There was not a positive association between the encounter of R. sanguineus and the infection for B. canis. The sequencing of two positive samples demonstrated close identity with B. canis vogeli already characterized in Brazil.
Mestre

The aim of this study was to analyze the epidemiological aspects of the canine babesiosis in animals of a city located on a semi-arid area of Minas Gerais state. Blood samples were randomly collected of Porteirinhas canine population - city situated in Itacambira region - based in a expected prevalence of about 20% and a confidence rate of about 90%, completing a total of 505 animals. Blood samples were collected in march2003 and were examined using the RIFI technique. On June, September and December of 2003, blood samples were collected in the same animals, except from those that were not found. At the same time, a form was filled with information about sex, breed and age of the dogs. The information was analyzed with the Windows Access, and was verified statistically with the Excel 8.0 and Epi-Info version 3.3. It was found a prevalence rate of antibodies anti-B. vogeli in about 18.81% dogs. Sex and age were not associated with seroprevalence for B. vogeli. Mongrels were significantly more positive than the dogs with defined breed. The incidence rate of canine babesiosis during the period of April to June (13.79%) was significantly higher than the incidence observed in the period of October to December (7.73%). Analysis of the data found on this study suggest that climatical conditions of the semi-arid areas interfere in the transmission of B. vogeli
O objetivo deste trabalho foi estudar os aspectos epidemiológicos da babesiose canina em animais de uma cidade localizada em área semi-árida do estado de Minas Gerais. Foi realizada uma amostragem aleatória na população canina da cidade de Porterinha, na região de Itacambira, baseada numa prevalência esperada de 20% e grau de confiança de 90%, perfazendo um total de 505 animais. Amostras de sangue foram coletadas no mês de março de 2003 e examinadas por meio da Reação de Imunofluorescência Indireta. Nos meses de junho, setembro e dezembro de 2003, amostras de sangue foram coletadas dos mesmos animais, excetuando-se aqueles que não foram localizados. Concomitante à coleta de sangue foi preenchido um formulário no qual constavam informações sobre o sexo, raça e idade do cão. Os dados foram armazenados no programa Windows Access, sendo analisados estatisticamente por meio do Excel 8.0 e Epi-info versão 3.3. Foi determinada uma prevalência de anticorpos anti-Babesia vogeli de 18,81%. Não foi encontrada relação entre os fatores idade e sexo com a soroprevalência para B. vogeli. A soroprevalência de B. vogeli em cães Sem Raça Definida foi significativamente maior em relação a animais de raça definida. A taxa de incidência da babesiose canina no período de abril-junho (13,79%) foi significativamente maior que a incidência observada de outubro-dezembro (7,73%). Análise dos dados obtidos neste trabalho sugerem que as condições climáticas das áreas semi-áridas interferem na transmissão da B. vogeli

La babésiose canine est une maladie parfois mortelle due a des protozoaires des genres Babesia et Theileria transmis par des tiques dont Rhipicephalus sanguineus. Malgré la présence soutenue de R. sanguineus dans le Sud de la France, son rôle dans la transmission de la maladie dans cette région reste incertain. D'autre part, le statut taxonomique de R. sanguineus reste confus et mérite d'être clarifié. Les objectifs de ce travail ont été de caractériser (i) les tiques R. sanguineus s.l. parasitant les chiens dans différentes régions du Sud de la France, (ii) les espèces des genres Babesia/Theileria potentiellement transmises par ces tiques et (iii) l'impact des variations génétiques et/ou des co-infections sur la compétence vectorielle. Au total 140 sangs de chien et 242 tiques R. sanguineus s.l. ont été récoltées et analysées entre 2010 et 2012 par PCR, PCR-RFLP et séquençage. De l'ADN de B. vogeli et B. canis a été détecté dans 13,6% et 12,9% des chiens et dans 10,7% et 1,7% des tiques respectivement avec, parfois, des prévalences significativement différentes entre les régions. Un seul chien a été détecté porteur d'ADN de T. annae. Des comparaisons phylogénétiques et phylogéographiques de séquences de R. sanguineus (ADNr-mt 12S et 16S) et B. vogeli (ARNr 18S) à l'échelle mondiale ont confirmé l'affiliation des tiques de France aux R. sanguineus des régions « tempérées » et suggèrent l'existence d'une coévolution entre le protozoaire et son vecteur. Ce travail apporte de nouvelles connaissances sur la babésiose canine en France et supporte l'existence d'au moins deux populations au sein du groupe sanguineus pouvant présenter des compétences vectorielles différentes
Study on Rhipicephalus sanguineus sensu lato ticks and their potential role in the transmission of canine babesiosis in France: prevalence of parasite infections, vector genetic diversity and epidemiology. Canine babesiosis is a potentially fatal disease caused by protozoa of the genera Babesia and Theileria transmitted by ticks including Rhipicephalus sanguineus. Although R. sanguineus is a major tick species in southern France, its role in the transmission of the disease in this area remain unknown. Moreover, the taxonomic status of R. sanguineus is controversial and needs to be clarified. The aims of this work were to characterize (i) the ticks R. sanguineus s.l. that parasitize dogs in different areas of southern France, (ii) the Babesia/Theileria species potentially transmitted by these ticks in this area and (iii) the impact of genetic variations and/or co-infections on vector competence. A total of 140 dog bloods and 588 R. sanguineus s.l. were collected from 2010 to 2012 from which all dogs bloods and 242 ticks were screened using specific PCR, PCRRFLP and sequencing. B. vogeli and B. canis DNA were evidenced in 13.6% and 12.9% of dogs and in 10.7% and 1.7% of R. sanguineus ticks respectively with sometimes significant prevalence differences between areas. A single case of T. annae infection was detected in a dog. Phylogenetic and phylogeographic analyzes performed at a global scale on 12S and 16S mt-rDNA for ticks and 18S rDNA for B. vogeli confirmed the affiliation of R. sanguineus from France with “temperate” species and suggested the existence of a coevolution process between the pathogen and its vector. This work provides new information on the epidemiology of canine babesiosis in France and supports the existence of at least two populations in the R. sanguineus group in the world with possible different vector competences

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Dentre as doenças emergentes causadas por artrópodes em cães destacam-se as infecções por agentes das espécies Leishmania chagasi, transmitida por flebotomíneos, Ehrlichia canis e Babesia canis transmitidas por carrapatos da espécie Rhipicephalus sanguineus. O presente estudo teve como objetivo realizar o diagnóstico sorológico e molecular de parasitos das espécies Leishmania chagasi, Ehrlichia canis e Babesia canis em amostras de soro e de baço, respectivamente. Foram utilizados 60 cães naturalmente infectados, eutanasiados no Centro de Controle de Zoonoses do Município de Campo Grande, Mato Grosso do Sul. Pela Reação de Imunofluorescência Indireta (RIFI), 60 (100%), 39 (65%) e 49 (81%) dos 60 animais amostrados foram sororeagentes frente aos antígenos de L.chagasi, E.canis e B.canis respectivamente. Também foi realizado o ensaio Imunoenzimático Indireto (ELISA-teste) para detecção de anticorpos da classe IgG anti-L. chagasi, onde sessenta cães (100%) mostraram-se soropositividade. Cinquenta e quatro cães (90%) mostraram-se positivos na reação em cadeia pela polimerase (PCR) para o gênero Leishmania sp e destes cinquenta e três animais, 53 (88%) foram positivos para o complexo L.Donovani. Vinte e sete animais (45%) foram positivos para Ehrlichia e destes animais todos foram positivos para E.canis pelo nested PCR e 2 (3,33%) cães foram PCR positivos para Babesia sp. Os amplicons foram confirmados por seqüenciamento e os DNAs mostraram 99% de similaridade genética para L.chagasi, E.canis e B.canis. Este estudo mostrou que os cães residentes em Campo Grande/MS, podem albergar mais de um hemoparasita, As co-infecções por agentes patogênicos intracelulares de células do sistema monocítico fagocitário, como é o caso da Ehrlichia e Leishmania, associadas ou não a Babesia...
Among emerging diseases caused by arthropods in dogs stand out infections by agents of the species Leishmania chagasi, transmitted by sandflies, Ehrlichia canis and Babesia canis transmitted by ticks of the species Rhipicephalus sanguineus. The present study aims to perform serological and molecular diagnosis of parasites of the species Leishmania chagasi, Ehrlichia canis and Babesia canis in samples of sera and spleen, respectively. A total of 60 naturally infected dogs were euthanized at the Zoonosis Control Center of the city of Campo Grande, Mato Grosso do Sul Through Immunofluorescence Assay (IFA), sixty (100%), 39 (65%) and 49 (81%) of the 60 animals sampled were reactive serum against the antigens L.chagasi, E.canis and B. canis, respectively. We also performed indirect immunosorbent assay (ELISA test) for detection of IgG antibodies anti-L. chagasi, where sixty dogs (100%) were seropositive. Fifty-four dogs (90%) were positive by PCR for the genus Leishmania, and of these fifty and three animals, 53 (88%) were positive for the complex L.Donovani. Twenty-seven dogs (45%) were positive for Ehrlichia sp and these animals were all positive for E.canis by nested PCR and two dogs (3%) were PCR positive for Babesia sp. The amplicons were confirmed by DNA sequencing and showed 99% genetic similarity to L.chagasi, E.canis and B. canis. This study showed that dogs living in Campo Grande / MS, can accommodate more than one haemoparasite. The co-infections by intracellular pathogens monocytic phagocyte system cells, as is the case with Ehrlichia and Leishmania, associated or not with Babesia (which multiplies in red blood cells), causing serious damage to the health of animals. The decline in physical condition and immune co-infections resulting from increases parasitemia, which in the case of agents transmitted by... (Complete abstract click electronic access below)

Orientador: Gilson Pereira de Oliveira
Coorientador: Gisele Braziliano de Andrade
Coorientador: Alvimar Jose da Costa
Banca: Carlos Noriyuki Kaneto
Banca: Heitor Miraglia Herrera
Resumo: Dentre as doenças emergentes causadas por artrópodes em cães destacam-se as infecções por agentes das espécies Leishmania chagasi, transmitida por flebotomíneos, Ehrlichia canis e Babesia canis transmitidas por carrapatos da espécie Rhipicephalus sanguineus. O presente estudo teve como objetivo realizar o diagnóstico sorológico e molecular de parasitos das espécies Leishmania chagasi, Ehrlichia canis e Babesia canis em amostras de soro e de baço, respectivamente. Foram utilizados 60 cães naturalmente infectados, eutanasiados no Centro de Controle de Zoonoses do Município de Campo Grande, Mato Grosso do Sul. Pela Reação de Imunofluorescência Indireta (RIFI), 60 (100%), 39 (65%) e 49 (81%) dos 60 animais amostrados foram sororeagentes frente aos antígenos de L.chagasi, E.canis e B.canis respectivamente. Também foi realizado o ensaio Imunoenzimático Indireto (ELISA-teste) para detecção de anticorpos da classe IgG anti-L. chagasi, onde sessenta cães (100%) mostraram-se soropositividade. Cinquenta e quatro cães (90%) mostraram-se positivos na reação em cadeia pela polimerase (PCR) para o gênero Leishmania sp e destes cinquenta e três animais, 53 (88%) foram positivos para o complexo L.Donovani. Vinte e sete animais (45%) foram positivos para Ehrlichia e destes animais todos foram positivos para E.canis pelo nested PCR e 2 (3,33%) cães foram PCR positivos para Babesia sp. Os amplicons foram confirmados por seqüenciamento e os DNAs mostraram 99% de similaridade genética para L.chagasi, E.canis e B.canis. Este estudo mostrou que os cães residentes em Campo Grande/MS, podem albergar mais de um hemoparasita, As co-infecções por agentes patogênicos intracelulares de células do sistema monocítico fagocitário, como é o caso da Ehrlichia e Leishmania, associadas ou não a Babesia... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Among emerging diseases caused by arthropods in dogs stand out infections by agents of the species Leishmania chagasi, transmitted by sandflies, Ehrlichia canis and Babesia canis transmitted by ticks of the species Rhipicephalus sanguineus. The present study aims to perform serological and molecular diagnosis of parasites of the species Leishmania chagasi, Ehrlichia canis and Babesia canis in samples of sera and spleen, respectively. A total of 60 naturally infected dogs were euthanized at the Zoonosis Control Center of the city of Campo Grande, Mato Grosso do Sul Through Immunofluorescence Assay (IFA), sixty (100%), 39 (65%) and 49 (81%) of the 60 animals sampled were reactive serum against the antigens L.chagasi, E.canis and B. canis, respectively. We also performed indirect immunosorbent assay (ELISA test) for detection of IgG antibodies anti-L. chagasi, where sixty dogs (100%) were seropositive. Fifty-four dogs (90%) were positive by PCR for the genus Leishmania, and of these fifty and three animals, 53 (88%) were positive for the complex L.Donovani. Twenty-seven dogs (45%) were positive for Ehrlichia sp and these animals were all positive for E.canis by nested PCR and two dogs (3%) were PCR positive for Babesia sp. The amplicons were confirmed by DNA sequencing and showed 99% genetic similarity to L.chagasi, E.canis and B. canis. This study showed that dogs living in Campo Grande / MS, can accommodate more than one haemoparasite. The co-infections by intracellular pathogens monocytic phagocyte system cells, as is the case with Ehrlichia and Leishmania, associated or not with Babesia (which multiplies in red blood cells), causing serious damage to the health of animals. The decline in physical condition and immune co-infections resulting from increases parasitemia, which in the case of agents transmitted by... (Complete abstract click electronic access below)
Mestre

碩士
中興大學
獸醫微生物學研究所
95
Laboratory diagnosis of canine babesiosis is based primarily on blood smear examination. However, visually distinguishing between Babesia gibsoni and Babesia canis is difficult because their sizes are very similar. Another way to distinguish between these two species is by DNA molecular techniques. DNA molecular techniques offer the advantages of sensititvity and specificity. We developed a PCR screening procedure to discriminate between these species based on differences in their 18S rRNA, hsp70 and P18 genes. In this screening procedure, our primers successfully amplify PCR products in B. gibsoni but not in B. canis. Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid method. It has high specificity, efficiency, and can be performed rapidly under isothermal condition. We determined that the sensitivities of the LAMP method and PCR method are equivalent, with detection limits of 10-6 each. A previously published study reported that B. gibsoni and B. canis could be distinguished from each other by LAMP within the 18S rRNA genes（Ikadai et al., 2004）. However, we report that in our hands, LAMP as previously published did not in fact distinguish these two species. Sequence analysis confirmed a 99% nucleotide identity within the 18S rRNA genes amplified by LAMP. Therefore, we designed primers for LAMP based on another gene, P18. This protocol successfully displayed specificity for detecting B. gibsoni.

碩士
國立陽明大學
寄生蟲學研究所
87
This study using specific monoclonal antibodies, which was desiged antigen capture enzyme-linked immunosorbent assay(ELISA), detection of circulating antigens of caine babesiosis cause by B. gibsoni. The monoclonal antibodies (Mabs) BG 1(IgG1 κ chain) , BG 2(IgG1 κ chain) and BG3 (IgM κ chain) , were prepared against merozoite antigens from 3 days’ culture of Babesia gibsoni . In the Western blotting, BG 1, BG 2 and BG 3 all can recognize antigen p63 from the supernatant of the culture medium. In addition, merozoite antigens p65 can be recognized by BG 1 and p170 can be recognized by BG 2 and BG 3. In the Indirect Fluorescent Antibody Test, however, the Mabs, BG 1, BG 2 and BG 3, can react with B. gibsoni in the dog’s blood or erythrocytes and showed the specific fluorescent reaction. An antigen capture enzyme-linked immunosorbent assay was designed with Mabs of BG 1, BG 2 and BG 3, to detect circulating antigens of B. gibsoni in blood of experimentally infected dogs. Meanwhile, blood smears from infected dogs were prepared for microscopic examination of B. gibsoni and served as control. The results showed that the antigens can be detected one to two days earlier by ELISA than that by direct blood smear.The Mabs did not react, in antigen capture ELISA or Western blots, with the common haemoparasites of dog, namely, B. canis, Ehrlichia platys, Dirofilaria immitis adult and larva, Schistosoma japonicum, Toxoplasma gondii and normal dog’s blood. Furthermore it is possible to detect circulating antigen in sera collected from dog during the acute or chronic phases of infection by using this test.

Background: Canine babesiosis is a tick-borne disease causing detrimental health effects on the domestic dogs with huge economic impact on the owners. The most complicated form of canine babesiosis is caused by a pathogenic Babesia rossi parasite. Canine babesiosis induced by B. rossi still remains the cause of mortality and morbidity in South African dogs, yet, the transcriptomic and genomic information of this parasite species is still not available. The transcriptomic and genomic information is essential in the disease development and processes for the design of effective disease control strategies. Consequently, our understanding of the mechanisms underlying the pathogenesis of the different genotypes of B. rossi remains limited. A previous study suggested a relationship between the parasite genotype and the disease phenotype. To date, thirteen B. rossi genotypes have been identified and associated with diverse clinical signs in their hosts. Hence the aim of this study was to sequence RNA from samples representing B. rossi genotypes, 19, 29 and 31, in order to have insight on the overall transcriptome of this parasite and to establish if there would be significant differences among the genotypes. Methodology: To screen for B. rossi positive samples, total DNA was extracted from 20 blood samples collected from sick domestic dogs presented at the Onderstepoort Veterinary Academic Hospital (OVAH). Babesia rossi infections were confirmed using the PCR-Reverse Line Blot (RLB) hybridization assay. Further confirmation of infection status was done by amplification of the B. rossi Erythrocyte Membrane Antigen 1 (BrEMA1) gene in all the DNA samples using qualitative PCR (qPCR), followed by sequencing of PCR products. Subsequently, total RNA was extracted from the 20 B. rossi-infected blood samples collected from the same dogs in which DNA was extracted. Three samples representing B. rossi genotypes 19, 29 and 31 were selected for transcriptome analysis. RNA sequencing was performed using the Illumina HiSeq 2000 to allow transcriptome analysis. De novo assembly was performed independently for all three transcriptomes using the Trinity software. The unigenes generated from specific transcriptome assemblies were subjected to global functional annotation using Blast2GO version 2.8.0 software, followed by KEGG database for annotation of biological pathways, and DAVID version 6.7, for COG classification to predict and classify their functions. Results: The sample representing B. rossi genotype 31 was excluded in the transcriptome analysis due to low RNA mass, which usually compromises the quality of the library used in RNA sequencing.Thus, a total of 26 747 238 and 25 709 627 paired-end reads were obtained from B. rossi genotypes 19 and 29, respectively. De novo transcriptome assembly produced a total of 3019 unigenes, with an average length of 419 bp and N50 of 362 bp in B. rossi genotype 19, and 2727 unigenes with an average length of 441 bp and N50 of 362 in B. rossi genotype 29. A total of 1193 unigenes were common between B. rossi genotype 19 and 29, while 1828 unigenes were exclusively detected in B. rossi genotype 19; and 1534 were specific to B. rossi genotype 29. Between the two B. rossi genotypes, a total of 4553 unigenes were obtained, representing the overall B. rossi transcriptome. From the overall transcriptome, 12.3% (n=558) of the unigenes could be annotated with 53 different gene ontology (GO) functional categories. About 34% (n=1550) of the unigenes represented in the overall transcriptome mapped to 237 KEGG pathways and only 2.5% (114) could be annotated in the COG database. Conclusion: Although, there were no striking differences in the transcriptomes of B. rossi genotypes 19 and 29, this study presents the first transcriptomic resource for B. rossi, which will highly contribute to our genetic understanding of B. rossi and provide a platform for future gene expression studies. Hypothetical proteins identified in this study will require further characterization as they may have a critical role in the biology and pathogenicity of B. rossi parasite.
Life and Consumer Sciences
M. Sc. (Life Sciences)

Dissertação de Mestrado Integrado em Medicina Veterinária, Ciências Veterinárias
A babesiose canina (ou piroplasmose) é causada por protozoários transmitidos por carraças e afeta cães em muitas partes do mundo. As duas principais espécies reconhecidas como agentes da doença são Babesia canis e Babesia gibsoni, que correspondem respetivamente a formas intraeritrocitárias (ou piroplasmas) grandes (3-5 μm) e pequenas (0,5-2,5 μm). As espécies B. canis e Babesia vogeli, anteriormente consideradas subespécies de B. canis, são os dois agentes da babesiose canina mais frequentemente assinalados na Europa, estando associados às carraças Dermacentor reticulatus e Rhipicephalus sanguineus, respetivamente. Os sinais clínicos da doença são variáveis e incluem essencialmente apatia, anorexia, perda de apetite, mucosas pálidas, icterícia, urina escura, epistaxis e dor. Outras manifestações clínicas incluem trombocitopenia e anemia hemolítica. A babesiose canina causada por piroplasmas grandes é endémica no Nordeste Transmontano, e tanto as espécies B. canis como B. vogeli já foram especificamente identificadas em cães desta área geográfica. O presente trabalho faz uma abordagem à babesiose canina no distrito de Bragança, focando essencialmente dados clínicos de animais em que foi diagnosticada a doença. Foram analisados os processos clínicos de 110 cães suspeitos de hemoparasitoses, tendo sido diagnosticada babesiose em 84 deles (76,4%) através de observação de piroplasmas grandes em esfregaço de sangue periférico. Destes 84 cães, 46 (45%) eram machos, 38 (55%) eram fêmeas, 44 (52,4%) tinham menos de 3 anos de idade e aproximadamente 70% (57/84) dos cães tinham aptidão para a caça. Em 39 cães (46,4%) verificou-se a presença de carraças. Os sinais clínicos mais frequentes foram apatia (76,1%; 64/84), pigmentúria (68,3%; 41/60), anorexia (50%; 42/84), mucosas anémicas (35,1%; 27/77) e icterícia (13%; 10/77). Setenta e três cães (87%) (54 do concelho de Bragança e 19 do concelho de Vinhais) habitavam dentro da área de delimitação do Parque de Montesinho-Nogueira. Os casos de babesiose canina deste estudo incidem predominantemente em dois períodos específicos, no outono (outubro, novembro e dezembro) e na primavera (março, abril e maio). Em 19 cães foi confirmada a presença de Babesia canis por PCR e posterior análise por sequenciação do gene 18S rARN. Confirma-se a existência de babesiose canina na área em estudo, o Nordeste Transmontano, o que deve ser levado em conta na clínica de pequenos animais em termos de diagnóstico e, sobretudo na aplicação de medidas profiláticas.
Canine babesiosis (or piroplasmosis) is a protozoal tick-borne disease with worldwide distribution. Two main species have been identified as aetiological agents of the disease in dogs: Babesia canis and Babesia gibsoni, which correspond to large (3–5 mm) and small (0.5–2,5 μm) intraerythrocytic parasitic forms (or piroplasms), respectively. Species B. canis and Babesia vogeli, previously considered as subspecies of B. canis, are the two most frequently reported agents of canine babesiosis in Europe, being associated with Dermacentor reticulatus and Rhipicephalus sanguineus ticks, respectively. The clinical manifestations of babesiosis are variable but essentially include lethargy, anorexia, decreased appetite, pale mucous membranes, jaundice, haemoglobinuria, epistaxis and pain. Thrombocytopenia and hemolytic anemia can also be present. Canine babesiosis caused by large piroplasms is endemic in the Northeast of Portugal and both species B. canis and B. vogeli have been molecularly identified in dogs from this geographical area. This dissertation presents an approach to the canine babesiosis in the district of Bragança, focusing essentially on clinical data from dogs diagnosed with babesiosis. From 110 dogs with suspected of a canine vector-borne disease, 84 were diagnosed with babesiosis (76.4%) by observing intraerythrocytic large piroplasms on peripheral blood smears. Of these 84 dogs, 46 (45%) were male, 38 (55%) were female, 44 (52.4%) had less than 3 years and approximately 70% (57/84) were hunting dogs. Thirty-nine dogs (46.4%) were found infested with ticks. The most frequent clinical signs were lethargy (76.1%, 64/84), haemoglobinuria (68.3%, 41/60), anorexia (50%, 42/84), pale mucous membranes (35.1%, 27/77) and jaundice (13%, 10/77). Seventy-three dogs (87%) (54 from the municipality of Bragança and 19 from the muncipality of Vinhais) lived within the limits of Montesinho-Nogueira Park. The present cases of canine babesiosis occurred mainly in two specific periods, during fall (October, November and December) and spring (March, April and May). In 19 of these dogs, the presence of B. canis was confirmed by polymerase chain reaction (PCR) and subsequent sequencing of the 18S rRNA gene. The presence of canine babesiosis in the Northeast of Portugal should alert small animal veterinary practitioners in terms of diagnosis and also to promote effective control measures against this vector-borne disease.

Dissertação de Mestrado Integrado em Medicina Veterinária
A babesiose ou piroplasmose canina é uma doença causada por hemoparasitas do género Babesia, que tem distribuição mundial e importante significado clínico. Os agentes do género Babesia são transmitidos por carraças da família Ixodidae e podem afetar diferentes espécies animais e seres humanos. No entanto, a maioria dos agentes não apresenta potencial zoonótico. O quadro clínico dos animais infetados é caracterizado por febre, letargia, anemia, trombocitopenia e hemoglobinúria. Pode manifestar-se de forma aguda ou crónica, e a gravidade da infeção por Babesia spp. pode variar desde infeção subclínica até doença fatal. O diagnóstico é estabelecido com base na análise da história e quadro clínico do animal, na identificação dos parasitas em esfregaços sanguíneos e através de testes serológicos e/ou moleculares positivos. O tratamento consiste na administração de fármacos antibióticos e antiprotozoários que eliminam os agentes. A babesiose canina é considerada uma parasitose emergente e em expansão para locais onde não é endémica, constituindo assim um problema de saúde cada vez maior e mais importante no mundo. Esta dissertação contém a descrição de três casos clínicos tratados no Hospital Veterinário do Baixo Vouga, na tentativa de abranger uma amostra de casos distintos de babesiose canina. Nesses três casos, o diagnóstico final foi estabelecido através da observação de formas parasitárias intra-eritrocitárias compatíveis com Babesia spp. em esfregaços sanguíneos. Foi possível conhecer a espécie de Babesia em dois dos casos com recurso a PCR e a sequenciação de ADN. O protocolo terapêutico aplicado consistiu na administração de dipropinato de imidocarb e doxiciclina, além do tratamento das complicações associadas à babesiose. Apesar de apresentarem manifestações clínicas e gravidade diferentes entre si, todos os animais tiveram recuperação clínica.
Canine babesiosis or piroplasmosis is caused by hemoparasites of the genus Babesia, which have a worldwide distribution and an important clinical significance. Piroplasms are transmitted by ticks of the Ixodidae family and can affect a variety of animal species, including humans; however, most agents do not have zoonotic significance. The clinical presentation of the infected animals is characterized by fever, lethargy, anemia, thrombocytopenia and hemoglobinuria. Babesiosis can be presented in an acute or chronic form, and severity can range from subclinical infection to fatal disease. The diagnosis is based on the clinical history and results of clinical exams, identification of parasites in blood smears and in positive results in serological and / or molecular tests. Treatment consists in administration of antibiotic and antiprotozoal drugs that eliminates the agents. Canine babesiosis is considered an emerging parasitosis, and is in expansion to areas previously considered as non-endemic, thus constituting an increasingly health problem in the world. This dissertation includes the description of three clinical cases treated at the Hospital Veterinário do Baixo Vouga, in an attempt to report different cases of canine babesiosis. The final diagnosis was established through identification of intra-eritrocytic inclusions compatible with Babesia spp. in blood smears. In two of the clinical cases, the species of Babesia was determined by PCR and DNA sequencing. The therapeutic protocol applied in all cases consisted in the administration of dipropinate imidocarb and doxycycline and support treatment. Despite the different clinical manifestations and severity, all animals had clinical recovery.

Dissertação de Mestrado Integrado em Medicina Veterinária apresentada à Universidade de Trás-os-Montes e Alto Douro
A babesiose ou piroplasmose é uma das doenças caninas com maior importância a nível mundial, sendo causada por parasitas intraeritrocitários do género Babesia. A sua transmissão dá-se por meio de ixodídeos infetados com os agentes patogénicos, que recebem a designação de piroplasmas. Na Europa, a região da bacia do Mediterrâneo é uma das mais afetadas. Em Portugal, a região do Nordeste Transmontano é a mais afetada, constituindo os cães de caça a maior parte dos casos. A ampla gama de manifestações clínicas depende muito das espécies de Babesia que causam a infeção e outros fatores que afetam a gravidade da doença, mas a maior parte dos sinais clínicos são comuns. Estes incluem febre, membranas mucosas pálidas, icterícia, vómito, hemoglobinúria, anorexia, perda de peso e fraqueza, entre outros. Os métodos de diagnóstico mais utilizados são a identificação direta de merozoítos (piroplasmas) nos eritrócitos no esfregaço de sangue periférico, métodos serológicos de deteção de anticorpos como o ELISA e amplificação de DNA. Há uma ampla gama de opções de tratamento, sendo que este passa pela administração de antiprotozoários e antibióticos para a eliminação dos agentes. Esta dissertação contém a descrição de alguns casos clínicos tratados dos CAMV Trás-os-Vet de Boticas e Montalegre, adaptando o conhecimento veterinário a um meio mais rural em que os meios podem ser limitados.
Babesiosis or piroplasmosis is one of the most important canine diseases worldwide and is caused by an intraerythrocyte parasite of the genus Babesia. The transmission occurs by ixodids infected with pathogenic agent, which is called the piroplasm. In Europe, the Mediterranean region is one of the most affected. In Portugal, the region of the Northeast of the Trás-os-Montes is the most affected, with the most part of cases being hunting dogs. The wide range of clinical manifestations depends largely on the species of Babesia which cause infection and other factors that affect the severity of the disease, however most clinical signs are common. These include fever, pale mucous membranes, anaemia, jaundice, vomiting, haemoglobinuria, anorexia, weight loss, weakness, among others. The most commonly used diagnostic methods are direct identification of merozoites in erythrocytes in the peripheral blood smear, serological methods for detecting antibodies such as ELISA and PCR amplification of DNA. There is a wide range of treatment options, which involves the administration of antiprotozoal agents and antibiotics for the elimination of the parasites. This dissertation includes the description of some clinical cases treated from the Trás-os-Vet veterinary medical centre of Boticas and of Montalegre, adapting veterinary knowledge to a rural environment where the means may be limited.

Dissertação de Mestrado Integrado em Medicina Veterinária
A babesiose é uma piroplasmose de importância mundial que afeta principalmente mamíferos. A doença é causada por um protozoário intracelular transmitido por vetores hematófagos. A apresentação da doença nos cães pode ser variada, desde uma doença inespecífica ligeira, até um quadro hiperagudo de colapso que leva ao óbito do paciente. Geralmente, os sinais clínicos observados são febre, anorexia, letargia e icterícia, acompanhados de anemia hemolítica, trombocitopenia, hemoglobinúria e bilirrubinúria. A presente dissertação inclui uma descrição de quinze casos clínicos de babesiose canina acompanhados durante o período de um ano e dois meses no Hospital Veterinário da Universidade de Trás-os-Montes (HV-UTAD). Os casos clínicos representam todos os animais nos quais foram detetadas estruturas compatíveis com Babesia spp. no esfregaço sanguíneo ou na deteção pela reação em cadeia da polimerase (PCR, do inglês “Polymerase Chain Reaction”), tivessem ou não outra infeção ou doença concomitante. Dos quinze casos que se apresentam só um deles foi confirmado por métodos nucleares (PCR). Uma série de fatores propiciam o aparecimento de Babesia spp. na região: as condições climáticas do Nordeste de Portugal afetam a distribuição das carraças vetor, a população canina errante e em contacto com a natureza é alvo destes parasitas, e as características intrínsecas dos detentores dos animais que descuidam os protocolos de desparasitação externa, bem como as condições de maneio.
Babesiosis is a worldwide-presenting piroplasmosis that mainly affects mammals. It originates from an intracellular protozoan parasite, which is transmitted by hematophagous vectors. The presentation of the disease in dogs can be varied, from a mild nonspecific disease, to a hyperacute collapse that leads to the death of the patient. Usually the clinical signs observed are fever, anorexia, lethargy and jaundice, accompanied by hemolytic anemia, thrombocytopenia, hemoglobinuria and bilirubinuria. The present dissertation includes a description of fifteen clinical cases of canine babesiosis accompanied during a one-year period at the Veterinary Hospital of the University of Trás-os-Montes, Vila Real. (HV-UTAD). Clinical cases represent all animals with structures compatible with Babesia spp. in the blood smear or in the detection with PCR, have or do not have another disease at the same time. Of the cases presented, only one was confirmed by nuclear methods (PCR). A series of factors favor the appearance of Babesia spp. in the region: the climatic conditions of the Northeast of Portugal affect the distribution of vector ticks, the roaming dog population and it’s contact with nature make them a target for these parasites, and the characteristics of the animal’s owners that neglect the protocols of external deworming and the conditions of handling.

Dissertação de Mestrado Integrado em Medicina Veterinária
A babesiose canina, também denominada de piroplasmose, trata-se de uma hemoparasitose provocada por um protozoário que apresenta tropismo para as células sanguíneas, mais especificamente para os eritrócitos. A transmissão deste agente etiológico ao seu hospedeiro ocorre pela sua inoculação através de artrópodes, sendo, neste caso, os ixodídeos os principais responsáveis. As espécies Babesia canis e Babesia gibsoni são responsáveis por provocar babesiose canina um pouco por todo o mundo, representado respetivamente formas intraeritrocitárias grandes (3-5 μm) e pequenas (0,5-2,5 μm). Anteriormente a primeira podia ser subdividida em 3 subespécies: Babesia canis canis, Babesia canis vogeli e Babesia canis rossi, sendo agora consideradas espécies individualmente. As espécies B. canis e B. vogeli são as mais frequentemente encontradas em Portugal e Europa em geral, e são transmitidas por vetores como o Dermacentor reticulatus e Rhipicephalus sanguineus respetivamente. A doença pode manifestar-se sob quatro formas: subclínica, aguda, hiperaguda e crónica, uma vez que a gravidade dos sinais clínicos está intimamente relacionada com o estado do hospedeiro (incluindo idade e condição imunitária) e com a espécie de Babesia envolvida. O tratamento da babesiose canina passa essencialmente pelo uso de anti-protozoários, mas acima de tudo, o mais importante baseia-se numa boa prevenção, através do uso de desparasitantes externos, administrados externamente ou per os. A presente dissertação inclui uma descrição de alguns casos clínicos acompanhados durante o período de estágio decorrido no Hospital Veterinário de Trás-os-Montes, Vila Real.
Canine babesiosis, sometimes referred to as piroplasmosis, is a hemoparasitosis caused by a protozoan that presents tropism to blood cells, more specifically to erythrocytes. The transmission of this etiological agent to its host occurs by its inoculation through vectors, being, in this case, the ixodídeos the main responsible ones. The species Babesia canis and Babesia gibsoni are responsible for causing canine babesiosis around the world, represented respectively large (3-5 μm) and small (0.5-2.5 μm) intraerocyte forms. Previously the first could be subdivided in 3 subspecies: Babesia canis canis, Babesia canis vogeli and Babesia canis rossi, being now considered species individually. The species Babesia canis and Babesia vogeli are most frequently found in Portugal and Europe in general and are transmitted by vectors such as Dermacentor reticulatus and Rhipicephalus sanguineus respectively. This disease can manifest itself in 4 forms: subclinical, acute, hyperacute and chronic, since the severity of the clinical signs is closely related to the state of the host (immune condition, age, etc.) and with the Babesia species involved. The treatment of canine babesiosis is essentially through the use of anti-protozoa, but above all, the most important is based on good prevention through the use of external parasites. This study refers and describe some case reports of dogs with clinical babesiosis that were observed during the pratice period at the Hospital Veterinário de Trás-os-Montes.

Orientação : Pedro Faísca ; co-orientação : Odete Almeida ; responsabilidade externa : Liliana Fernandes
As doenças transmitidas por vectores ocorrem pela picada, nomeadamente de mosquitos, flebótomos, ixodídeos e pulgas podem ser causadas por parasitas, bactérias ou vírus. Para além de algumas apresentarem prevalências elevadas e/ou serem endémicas, têm importância acrescida por muitas serem zoonoses. Nos últimos anos, tem havido uma emergência ou reemergência de algumas doenças, e muitas associadas a alterações epidemiológicas, sobretudo na sua expansão geográfica. Vários factores afectaram este fenómeno, especialmente o aumento da temperatura, que leva à alteração do microclima, biologia e comportamentos alimentares dos vectores, bem como as viagens de animais para zonas endémicas e a sua importação destas zonas. O objectivo do presente estudo foi conhecer e caracterizar a existência de doenças transmitidas por vectores no cão, numa clínica veterinária em Cascais, onde foram recolhidos dados retrospectivamente de 1 de Janeiro de 2010 e 29 de Fevereiro de 2012, e relacionou-se o seu diagnóstico com possíveis factores de risco, como a idade, a época do ano, sexo e raça. A amostra foi obtida através da base de dados da clínica, tendo sido analisados 42 cães com suspeita de Babesiose, 43 de Ehrlichiose, 46 de Rickettsiose e 190 de Leishmaniose. A Leishmaniose Canina foi a infecção com maior número de casos positivos, 53 em 190 (28%), seguida da Rickettsiose Canina com 40 em 46 (87%). A Babesiose apresentou apenas 8 casos em 42 (19%) e Ehrlichiose com 4 em 43 (9%). Foram também verificadas co-infecções entre Rickettsia conorii e Ehrlichia canis (n=4), Rickettsia conorii e Babesia canis (n=8) e Leishmania infantum e Rickettsia conorii (n=2). Os factores analisados não revelaram diferenças estatisticamente significativas no diagnóstico destes agentes.
Vector borne diseases, caused by parasites, bacterias or viruses can be transmitted by mosquitoes, sandflies, fleas and tick bites. The prevalence of these diseases is increasing, and some of them have a zoonotic character, which strengthen their importance. The aim of this study was to determine and characterize vector borne diseases in dogs, at a Veterinary Clinic located in Cascais (Portugal). A retrospective study was conducted, comparing vector borne diseases diagnosis with possible risk factors, such as age, time of year, sex and breed, in that particular geographic location, during the period of 1 January 2010 to 29 February 2012. There were a total of 207 patients of dogs who presented signs compatible with vector borne diseases: 42 suspected of Babesiosis, 43 suspected of Ehrlichiosis, 46 canine Ricketsiosis and 190 suspected of Leishmaniosis. It was concluded that in the area covered by this study, the elected factors didn’t have any statistic correlation with the positive diagnosis. It was also determined that Leishmaniasis had a bigger onset of positive cases, 53 out of 190 (28%), Ricketsiosis had 40 out 46 (87%), Babesiosis, 8 out of 42, (19%) and Ehrlichiosis, 4 out of 43, 9%.